应用竞争PCR检测丙酸盐对体外培养牛肝细胞丙酮酸羧化酶mRNA水平的影响.PDFVIP

See discussions, stats, and author profiles for this publication at: /publication/242280587 Effection of Propionate on the Expression of Pyruvate Carboxylase mRNA in vitro culture bovine Hepatocyte Determinated by... Article C TAT ONS READS 0 14 6 authors, including: Xia Cheng Wang Zhe Heilongjiang Bayi Agricultural University UNSW Sydney 52 PUBL CAT ONS 156 C TAT ONS 4 PUBL CAT ONS 1 C TAT ON SEE PROF LE SEE PROF LE Some of the authors of this publication are also working on these related projects: Nutritional metabolic diseases of dairy cows View project All content following this page was uploaded by Xia Cheng on 16 August 2015. The user has requested enhancement of the downloaded file. 应用竞争PCR检测丙酸盐对体外培养牛肝细胞丙酮酸羧化酶 mRNA 水平的影响 徐闯，夏成，刘国文，王哲*，李家奎，张乃生，王兴龙，欧阳红生，张永亮 （吉林大学农学部畜牧兽医学院，吉林 长春 130062 ） 摘 要：应用 PC(丙酮酸羟化酶)基因组与 PCcDNA 相比包含内含子序列及在其中插入外源 DNA 序列的方 法，改变 PCcDNA 的长度，成功构建 PCcDNA 的竞争 DNA 模板。然后应用竞争 PCR 方法研究丙酸盐对 体外培养新生牛单层肝细胞 PC mRNA 水平的影响。使单层肝细胞培养液中丙酸钠浓度分别为 0 mmol/L、 1.5 mmol/L、2.5 mmol/L 、3.5 mmol/L、4.5 mmol/L 、8.5 mmol/L 和 11.5 mmol/L，处理24 小时，提总 RNA 、 逆转录，在同一体系中用相同引物扩增目的带和竞争模板带。结果表明：随着丙酸钠浓度的升高 PC mRNA 水平呈上升趋势。指示肝细胞内 PCmRNA 的表达水平受培养液中丙酸钠浓度的影响。 关键词：丙酮酸羧化酶mRNA ；竞争PCR ；肝细胞；内含子 Effection of Propionate on the Expression of Pyruvate Carboxylase mRNA in vitro culture bovine Hepatocyte Determinated by Competive Reverse Transcription Polymerase Chain Reaction XU Chuang, XIA Cheng, LIU Guo-wen, WANG Zhe*, LI Jia-kui, ZHANG Nai-sheng, WANG Xing-long, OU YANG Hong-sheng, ZHANG Yong-liang （Agriculture Departme